WO2012135237A2 - Procédés, procédures et systèmes commerciaux pour la collecte, le stockage cryogénique et la distribution de formules cosmétiques à partir d'une matière biologique obtenue à base de cellules souches - Google Patents

Procédés, procédures et systèmes commerciaux pour la collecte, le stockage cryogénique et la distribution de formules cosmétiques à partir d'une matière biologique obtenue à base de cellules souches Download PDF

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Publication number
WO2012135237A2
WO2012135237A2 PCT/US2012/030774 US2012030774W WO2012135237A2 WO 2012135237 A2 WO2012135237 A2 WO 2012135237A2 US 2012030774 W US2012030774 W US 2012030774W WO 2012135237 A2 WO2012135237 A2 WO 2012135237A2
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WO
WIPO (PCT)
Prior art keywords
product
stem cell
cosmetic
biological
biological sample
Prior art date
Application number
PCT/US2012/030774
Other languages
English (en)
Other versions
WO2012135237A3 (fr
Inventor
John S. Arnone
Ensley BURT
Original Assignee
Arnone John S
Burt Ensley
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Arnone John S, Burt Ensley filed Critical Arnone John S
Priority to US14/008,024 priority Critical patent/US11389394B2/en
Priority to JP2014502724A priority patent/JP2014518846A/ja
Priority to CN201280026116.1A priority patent/CN104662532B/zh
Publication of WO2012135237A2 publication Critical patent/WO2012135237A2/fr
Publication of WO2012135237A3 publication Critical patent/WO2012135237A3/fr
Priority to HK15110032.2A priority patent/HK1209503A1/xx
Priority to US17/844,198 priority patent/US20220323342A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06Q—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES; SYSTEMS OR METHODS SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES, NOT OTHERWISE PROVIDED FOR
    • G06Q50/00—Systems or methods specially adapted for specific business sectors, e.g. utilities or tourism
    • G06Q50/10—Services
    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16H—HEALTHCARE INFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR THE HANDLING OR PROCESSING OF MEDICAL OR HEALTHCARE DATA
    • G16H10/00—ICT specially adapted for the handling or processing of patient-related medical or healthcare data
    • G16H10/40—ICT specially adapted for the handling or processing of patient-related medical or healthcare data for data related to laboratory analysis, e.g. patient specimen analysis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/10—General cosmetic use
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying

Definitions

  • the invention is directed to a business method for collection, cryogenic storage and distribution of cosmetic formulations from an obtained stem cell based biological sample material.
  • the invention is directed to a business method for the processing and production of a stem cell based cosmetic.
  • the method is initiated by designating a stem cell based biological material and thereafter, collecting a premium for defined services for collection, processing and transportation of the stem cell based biological material are established.
  • the business method continues by obtaining the stem cell based biological material from a source and processing the stem cell based biological material to form at least one biological product for use in a cosmetic product. Testing the at least one biological product for quality control will be performed at least at the completion of the "processing" and prior to moving forward with producing any cosmetic product.
  • the invention is directed to a system for producing a stem cell based cosmetic formulation product.
  • the system includes a stem cell based biological material collection system and a stem cell based biological material transportation system for transporting the components of the stem cell based biological material collection system to a processing facility.
  • the processing facility has a database which processes the stem cell based biological material to form at least one biological product.
  • a storage facility stores the at least one biological product and a retrieval system is included for distributing the at least one biological product.
  • a cosmetic formulation processing facility prepares a cosmetic formulation based on the at least one biological product
  • the invention is directed to a method for preparing an autologous cosmetic composition including the steps of obtaining a biological sample from a client and separating the biological sample to obtain a defined separation product.
  • the defined separation product is combined with a growth media to obtain stem cells grown to confluence.
  • the stem cells grown to confluence are combined with a non-growth media to obtain an active substance which is isolated.
  • the isolated active substance is combined with a cosmetic formulation.
  • the invention is directed to a method of preparing components for a stem cell based cosmetic by ultrasonic cavitation including the steps of forming a stromal vascular fraction by ultrasonic cavitation of an adipose tissue sample and thereafter, combining the stromal vascular fraction with a growth media to grow stem cells to confluence.
  • the stem cells are isolated and thereafter combined with a non-growth media allowing secretion of cytokines from the stem cell to produce an active substance, wherein the active substance is combined with a cosmetic formulation to form a cosmetic product.
  • the invention is directed to a method to prepare an ethnic stem cell based cosmetic product including the steps of obtaining a biological sample from a donor of a defined ethnicity and processing the biological sample to obtain an active substance.
  • the active substance is combined with a cosmetic formulation.
  • the invention is directed to a business method for continuous supply of an ethnic stem cell based cosmetic product.
  • the business method of the present embodiment is initiated by selecting a donor of a defined ethnicity and obtaining a biological sample from the donor of a defined ethnicity. Coordinating the processing of the biological sample, wherein the biological sample is disassociated to obtain a defined separation product and thereafter, processing the defined separation product forms an active substance.
  • the active substance is combined with a cosmetic formulation that contains an active ingredient for a defined desired property of an ethnic group to form an ethnic stem cell based cosmetic product.
  • the ethnic stem cell based cosmetic product is commercially distributed to consumers in various methods recognized in the industry.
  • FIG 1 is a schematic view of the system of the present invention.
  • FIG 2 is a flowchart illustrating the process of preparing a cosmetic product via two methods of separation, achieving a defined separation product to obtain an active substance for forming a cosmetic product;
  • Fig. 3 is a flowchart illustrating the business method for continuous supply of an ethnic stem cell based cosmetic product.
  • the invention is directed to a business method for the processing and production of a stem cell based cosmetic.
  • the method is initiated by designating a stem cell based biological material.
  • the source of the stem cell based biological material could vary to include an autologous client, a homologous source or a commercially obtained composition, e.g. a medium composition which includes stem cells.
  • the commercially obtained composition could be from a heterologous source, most commonly bovine.
  • the business method of the present invention includes defined premium collecting methods and information recordation designed for particular services based on the stem cell based biological material source (as discussed in detail in the examples herein).
  • the business method continues by obtaining the stem cell based biological material from a source as previously discussed. It will be appreciated that methods to obtain the stem cell based biological material will reflect standard medical services based on autologous, homologous or heterologous stem cells which form the basis of a cosmetic product.
  • Processing the stem cell based biological material to form at least one biological product for use in the cosmetic product is defined by the requirements of the stem cell based biological material and client information tracking, in addition to, and separate from, "digestion” protocols.
  • client information tracking in addition to, and separate from, "digestion” protocols.
  • One of skill in the art will recognize that various methods, manual, mechanical and/or chemical can be used for digestion.
  • the particular process and information obtained will depend on the source of the stem cell based biological material.
  • autologous samples obtained will require the greatest amount of information "tracking" in addition to the probability of the increased “digestion” steps.
  • commercially obtained (heterologous) materials will require the least amount of information tracking and "digestion”.
  • Testing the biological products for quality control will be performed at least at the completion of the "processing" and prior to moving forward with producing any cosmetic product. All testing will be at a minimum as required by the United States Food and Drug Administration.
  • a vehicle formulation is prepared for the particular cosmetic product desired.
  • the vehicle formulation will be based on 1) the "defined” biological product, and 2) the requirements of the particular cosmetic product desired.
  • One of skill in the art will appreciate that a vehicle formulation for a "foot cream” will be very different from an "eye serum”.
  • the formulation will coordinate with information obtained from each step of the business method of the present invention to establish a formulation which will efficiently and effectively allow the expression of all the components of the biological product.
  • the biological product is combined with the vehicle formulation for the particular cosmetic product to obtain the particular cosmetic product desired.
  • Core to this business method is establishing a method of combination which allows exponential benefits of the active ingredients and is cost effective based on the source of the stem cell based biological material.
  • the business method may further include coordinating a cosmetic procedure by a service provider using the particular cosmetic product.
  • a service provider most commonly a physician or trained technician.
  • the continued relationship will allow the cosmetic formulation to be applied by the service provider during additional sessions and "check-ups".
  • Coordinating the application of the particular cosmetic product with services most commonly dermatological services such as peelings, facials etc., will allow the client the confidence that the particular cosmetic product is being used and applied correctly, and most importantly, most effectively.
  • Having the opportunity to have the particular cosmetic product applied by a service provider in coordination with other treatments, when the particular cosmetic product can be most effective will build the relationship between the client and service provider to allow a long term relationship to obtain the results the client desires.
  • the business method may include cryopreserving the at least one biological product so that once obtained it can be used for future cosmetic preparations and allows the same at least one biological product to be used for different "particular" cosmetic products.
  • the business method can be repeated upon the quality control testing allowing re-ordering by a client to obtain a particular cosmetic product from the at least one biological product.
  • Example 1 is directed to a business method for collection, cryogenic storage and distribution of an autologous biological sample material to prepare a particular cosmetic product.
  • the method is initiated by collecting a premium for defined services for collection, cryogenic storage and distribution of a biological sample material and thereafter coordinating the collection of a biological sample of a customer by (i) paying a predetermined fee in support of physician services performed for collection of the biological sample and (ii) supplying a collection system including a plurality of components for collection and transportation of the biological sample.
  • This initial part of the business method is important not only to obtain the sample but to initiate the business relationship of the customer and business entity.
  • the customer, physician and business entity will gain an understanding of the "big picture" and long-term relationship of this collaboration so as to appreciate the benefits, rights, obligations and costs (as explained herein).
  • the pre-determined fee for a physician to obtain the biological sample could vary but will , mostly likely be limited to costs relating to the collection system and transportation to the processing facility. However, the cost will be a one-time set fee which will be agreed upon by the client before initiating the procedure to obtain the sample.
  • the collection system is a defined set of components which are designed for coordination of the business method.
  • the collection system includes an identification material for the obtained biological sample. This is most commonly a defined group of standard forms which may include coded labels for use with an encoded program (as discussed herein).
  • Client sample bags include the same coded labels for use with the encoded program. These labels will comply with state and federal regulations, e.g. 21 CFR 1 1 .
  • the collection system further includes a transportation box which may be commercially manufactured and coordinated with a transportation carrier, e.g. Biologies Box by FedEx. Transportation labeling will also include the same coded labels for use with the same encoded program; in addition to information regarding shipment location.
  • a transportation carrier e.g. Biologies Box by FedEx.
  • Transportation labeling will also include the same coded labels for use with the same encoded program; in addition to information regarding shipment location.
  • the method continues by obtaining the biological sample from the client and transporting the biological sample in the collection system to a processing facility.
  • the collection system components are introduced to a processing module of a database via a log-in port; having the encoded program.
  • the database will be custom-designed to process and store "eProtected" health information using a commercially available program such as Microsoft's Access program.
  • the database will include but is not limited to, the information obtained from the collection system to coordinate the "client sample with the client”; such as the information included in the patient-specific bar-coded client sample bags. This information will also be included in a standardized form.
  • the database will be searchable and may be programmed to produce all the various forms associated with this process.
  • the autologous biological sample is processed by digestion to separate and isolate material. Isolating the biological sample and ensuring the quality of the biological sample is an imperative key to the commercial success of the method. Testing is performed for quality control of the isolated material for cryopreservation (if requested) and thereafter, the isolated material is cryopreserved in at least one aliquot and stored.
  • the isolated material Upon request for use the isolated material is thawed in the at least one aliquot. The isolated material is then prepared into at least one cosmetic autologous formulation including the isolated material in the at least aliquot.
  • the client can "re-order" from a cryopreserved sample as desired until the sample is exhausted. It will be appreciated that additional products can be obtained by repeating this process initiating with obtaining the autologous sample.
  • the business method further comprises coordinating a cosmetic procedure by a service provider using the at least one cosmetic autologous formulation.
  • a cosmetic procedure by a service provider using the at least one cosmetic autologous formulation.
  • the cosmetic autologous formulation is formulated for a defined section of the body, including but not limited to a face cream, foot cream or eye cream.
  • the obtained biological sample is an adipose tissue sample, wherein the isolated material in the form of a stem cell pellet consisting essentially of a mixture of pre-adipocytes, adipose-derived mesenchymal stem cells, microvascular endothelial cells, endothelial progenitor cells, and monocytes.
  • the invention is directed to a business method for the processing and production of a stem cell based cosmetic.
  • the method is initiated by designating a stem cell source from a heterogeneous source.
  • the source of the stem cells is a commercially obtained composition, e.g. a medium composition which includes stem cells.
  • the commercially obtained composition could be from a heterologous source, most commonly bovine.
  • a fee is collected to cover the costs of obtaining the commercial composition, processing, quality testing and preparation of the formulation and finally, preparation of particular cosmetic product.
  • the heterologous stem cell composition is obtained which form the basis cosmetic product.
  • Processing the composition to form at least one biological product for use in the cosmetic product includes requirements of stem cells sample and client information tracking in additions and separate from "digestion" protocols. Additionally, any process or digestion which is required by the manufacturer of the commercial product will be preformed based on the manufacturer instructions, if any. In any process, the commercial product will be digested so as to be acceptable for combination with a vehicle formulation. This includes cryopreservation procedures, if required. It will be appreciated that cryopreservation may not be required or desired.
  • Testing the biological products for quality control will be performed at least at the completion of the "processing" and prior to moving forward with producing any cosmetic product. All testing will be at a minimum as required by the United States Food and Drug Administration.
  • a vehicle formulation is prepared for the particular cosmetic product desired.
  • the vehicle formulation will be based on 1) the "defined” biological product, and 2) the requirements of the particular cosmetic product desired.
  • One of skill in the art will appreciate that a vehicle formulation for a "foot cream” will be very different from an "eye serum”.
  • the formulation will coordinate with information obtained from each step of the business method of the present invention to establish a formulation which will efficiently and effectively allow the expression of all the components of the biological product.
  • the biological product is combined with the vehicle formulation for the particular cosmetic product to obtain the particular cosmetic product.
  • vehicle formulation for the particular cosmetic product
  • various methods, manual, mechanical and/or chemical can be used to combine the vehicle formulation and biological product. Core to this business method is establishing a method of combination which allows exponential benefits of the active ingredients and is cost effective based on the source of the stem cells.
  • the business method may further include coordinating a cosmetic procedure by a service provider using the particular cosmetic product, as discussed herein.
  • the invention is directed to a system for producing a stem cell based cosmetic formulation product 10.
  • the system 10 includes a stem cell based biological material collection system 12 and a stem cell based biological material transportation system 14 to a processing facility 16, having a database 18.
  • the database 18 includes an encoded program 20 to organize and store information regarding the stem cell based biological material and recording information. This includes information on standardized forms 22 which can be used in various parts of the system 10.
  • the collection system 12 has been defined in the previous embodiments and is usually in a "fitted box" for ease in use.
  • the collection system transportation system 14 is usually in coordination with a commercial carrier, such as Fed Ex®, which has the ability to transport medical samples using specific equipment in compliance with local, state and federal regulations.
  • the at least one biological product is located in a storage facility 24.
  • This is commonly a "freezer” of a defined size, having the capability and construction to coordinate with a second (permanent) storage facility (not shown) such as ovare® Bio-Logistics.
  • a strategic partnership is formed within the business method of the current invention.
  • the processed at least one biological product does not require cryopreservation and can be immediately used to form the particular cosmetic product.
  • the system 10 may not require a storage facility wherein no cryopreservation is required. In situations, wherein no cryopreservation is desired, the storage facility could be limited to an acceptable biological container at a preservation temperature.
  • the system 10 may include a retrieval system 26 for distributing the at least one biological product to multiple "end-user" opportunities (including the Donor as illustrated in Fig. 1 ).
  • the initial fees obtained are limited to the collection, digestion and initial storage of the stem cell based biological material and at least one biological product but does not include fees for the preparation of the cosmetic formulation.
  • a cosmetic formulation processing facility 28 prepares a cosmetic formulation based on the at least one biological product obtained.
  • the cosmetic formulation processing facility 28 accepts the "thawed" isolated material or material which has not been cryopreserved.
  • the cosmetic formulation can be multiple products including but not limited to face, eye and /or foot creams or serum.
  • the invention is directed to a method for preparing an autologous cosmetic composition including the steps of obtaining a biological sample from a client and separating the biological sample to obtain a defined separation product.
  • the defined separation product is combined with a growth media to obtain stem cells grown to confluence.
  • the stem cells grown to confluence are combined with a non-growth media to obtain an active substance which is isolated.
  • the isolated active substance is combined with a cosmetic formulation.
  • the combination of the isolated active substance and the cosmetic formulation creates a desired cosmetic product.
  • the biological sample is adipose tissue, obtained by aspiration 1 12; most commonly by a medical professional.
  • the invention is not limited to adipose tissue but may be directed to other biological samples, e.g. platelet rich plasma (PRP).
  • PRP platelet rich plasma
  • the defined separation product in the present embodiment is a stromal vascular fraction (SVF); the portion of adipose tissue that supports growth. More particularly, SVF is the lipoaspirate obtained from liposuction minus the fat cells (adipocytes). Apart from adipocytes, the SVF contains a variety of other cells such as pre-adipocytes, endothelial cells, smooth muscle cells, pericytes, fibroblasts, and adult stem cells (ASCs). In addition, the SVF also contains blood cells from the capillaries supplying the fat cells.
  • SVF stromal vascular fraction
  • SVF in addition to the adipocyte endocrine secretions, also contains growth factors such as transforming growth factor beta (TGF), platelet-derived growth factor (PDGF), and fibroblast growth factor (FGF), among others. This is consistent with the secretions of cells in the presence of an extracellular matrix.
  • TGF transforming growth factor beta
  • PDGF platelet-derived growth factor
  • FGF fibroblast growth factor
  • separation of the adipose tissue can be achieved by ultrasonic cavitation, primarily associated with cell disruption (lysis) or disintegration.
  • ultrasonic cavitation when sonicating liquids at high intensities, the sound waves that propagate into the liquid media result in alternating high-pressure (compression) and low-pressure (rarefaction) cycles, with rates depending on the frequency.
  • high-intensity ultrasonic waves create small vacuum bubbles or voids in the liquid.
  • separation can be achieved by enzymatic digestion of the adipose tissue 1 14B, wherein chemical substances separate the cell to obtain the defined separation product.
  • the stromal vascular fraction is combined with a growth media to grow stem cells to confluence 1 16.
  • the growth media can be of a variety of commercially available products having the composition to allow growth. However, specie specific media can be used which will eliminate any type immunological reaction, for example, the media described in U.S. Patent No. 7,989,205. One of skill in the art would recognize this step of the process can be repeated to obtain a defined or required number of cells.
  • an enzyme e.g. Tripson
  • an enzyme can be added to digest the linkage from "the surface” of the container in which "growth occurred", wherein the stem cells "float” in the liquid media.
  • the stem cells are isolated by pouring off the growth media and are combined with a non-growth media 1 18.
  • the non-growth media for example media limited to carbohydrates, is added to the container in which growth occurred. This allows secretion of cytokines from the stem cells into the "non-growth" media to produce the active substance.
  • the active substance is combined with a cosmetic formulation 120 having (i) a ph of between 6.0 and 8.0, and (ii) contains no proteases and (iii) is sterile.
  • the present embodiment further appreciates including in the defined separation product and growth media combination, tropoelastin to form the active substance. Adding tropoelastin when combining the growth media and the stromal vascular fraction will add additional properties to the active substance.
  • An autologous cosmetic product is formed by the method having defined properties for multiple uses.
  • the autologous cosmetic product achieved by the method of the present embodiment can serve multiple purposes including regenerative, anti-aging and beauty enhancement products but also to work with the body in multiple capacities to achieve designated ends.
  • Such capacities would include a method for stimulating skin (dermis) cells by contacting the dermis of a living individual with an autologous cosmetic product of the present embodiment.
  • the invention is directed to a method of preparing components for a stem cell based cosmetic by ultrasonic cavitation.
  • ultrasonic cavitation in a defined method for preparation of a cosmetic, focuses the properties and advantages of this method for preparation of components to achieve a specific objective; components which have properties which will allow preparation of the cosmetic.
  • ultrasonic cavitation is used to obtain components (via the separation activity) which "dovetail" with the other "steps" of the method for preparing an autologous cosmetic composition presented herein or related method for the same purpose.
  • Ultrasonication is an effective means to break cell structures. This effect can be used for the extraction of intracellular materials, e.g. starch from the cell matrix.
  • Ultrasonication generates alternating high-pressure and low-pressure waves in the exposed liquid.
  • the ultrasonic waves create small vacuum bubbles in the liquid that collapse violently during a high-pressure cycle; termed "cavitation".
  • the implosion of the cavitation bubble causes strong hydrodynamic shear-forces.
  • the shear forces can disintegrate fibrous, cellulosic material into fine particles breaking the walls of the cell structure.
  • the cell wall material is being broken into small debris. This effect can be used for fermentation, digestion and other conversion processes of organic matter.
  • ultrasonication makes more of the intra-cellular material e.g. starch as well as the cell wall debris available to the enzymes that convert starch into sugars. More importantly to the present embodiment, ultrasonic cavitation increases the surface area exposed to the enzymes during liquefaction or saccharification.
  • the cell yield by ultrasonic cavitation requires a high concentration of serum for growth of the cells for use in obtaining a cosmetic product. Without recognition of this requirement, the yield of cell growth does not form a basis for use in a cosmetic product nor does it allow for a successful commercial method.
  • the method of the present embodiment includes the steps of forming a stromal vascular fraction by ultrasonic cavitation of an adipose tissue sample in a defined manner to achieve a successful defined separation product which can be used within a defined method for making a cosmetic product.
  • the resultant stromal vascular fraction is combined with a growth media to grow stem cells to confluence.
  • the stem cells are isolated and thereafter combined with a non-growth media allowing secretion of cytokines, and other growth materials, from the stem cell to produce an active substance, wherein the active substance is combined with a cosmetic formulation to form a cosmetic product.
  • the active substance is combined with a cosmetic formulation having (i) a ph of between 6.0 and 8.0, and (ii) contains no proteases and (iii) is sterile and further, the growth media is either a commercially available media or a specie specific media. Additionally, as in the previous embodiment, tropoelastin can be combined with the growth media and stromal vascular fraction in obtaining the active substance.
  • the autologous cosmetic product formed by the method will have properties commensurate with components obtained by ultrasonic cavitation.
  • the invention is directed to a method to prepare an ethnic stem cell based cosmetic product including the steps of obtaining a biological sample from a donor of a defined ethnicity and processing the biological sample to obtain an active substance.
  • the donor is most commonly a person having superior qualities in regard to physical appearance including facial features, muscular appearance and skin quality.
  • the stem cell donor will exemplify the ethnic characteristic desired by consumers of cosmetic products.
  • the active substance is combined with a cosmetic formulation.
  • the cosmetic formulation in the present embodiment contains an active ingredient for a defined desired property of an ethnic group.
  • the formulation will be complementary to the active ingredient so as to allow a potential synergistic relationship but will also contain components which will have desired results of a particular ethnic group of the donor, e.g. a skin lightener.
  • Processing the biological sample includes separating the biological sample to obtain a defined separation product.
  • the defined separation product is most commonly an SVF obtained as discussed herein and combined with a growth media to obtain stem cells grown to confluence.
  • the stem cells grown to confluence are isolated and thereafter, combined with a non-growth media to obtain an active substance, which is isolated.
  • the active substance is combined with a cosmetic formulation as described herein to form an ethnic stem cell based cosmetic.
  • the invention is directed to a business method for continuous supply of an ethnic stem cell based cosmetic product 200.
  • the business method of the present embodiment is initiated by selecting a donor of a defined ethnicity 210 and obtaining a biological sample from the donor of a defined ethnicity 212. As discussed in the previous embodiment, the donor selected will illustrate the desired characteristics associated with an ethnic group.
  • the processing of the biological sample is coordinated, wherein the biological sample is disassociated to obtain a defined separation product and thereafter processing the defined separation product forms an active substance 214.
  • the coordination and processing can be achieved in defined manners which will contribute to the final consumer cosmetic product as discussed herein.
  • the active substance is combined with a cosmetic formulation that contains an active ingredient for a defined desired property of an ethnic group to form an ethnic stem cell based cosmetic product 216.
  • the active ingredient and formulation will be selected so as to satisfy the desired properties of the ethnic stem cell based cosmetic product but also in a manner which is cost effective for commercial success.
  • the ethnic stem cell based cosmetic product is commercially distributed to a plurality of customers in various methods recognized in the industry 218.
  • the method is initiated by designating an autologous stem cell based biological material of adipose tissue from an adult male.
  • a premium is collected for defined services for collection, transportation, processing and distribution of the stem cell based biological material.
  • the particular collection protocols are defined, coordinated and implemented based on the services required.
  • a shipment package including a defined client sample container is forwarded to the location of extraction of the biological sample.
  • the "shipment package” or “collection system” includes various components in addition to the defined container with the client information for extraction of the client/adipose tissue sample by a physician.
  • an additional 60mL syringe was included to the collection system in this example.
  • the instructions are amended to have the physician load both syringes with lipoaspirate, let stand 10 minutes then inject both into one collection bag of the collection system.
  • Inspecting the shipment package includes ensuring that (a) the defined client container is not past a defined expiration date, (b) the client sample was collected within the past 48 hours, and (c) the recording information is accurate. If the conditions of a, b, or c are not met, then the sample is not acceptable and must be discarded as biohazard waste. This discard will be recorded for organization and "tracking" of the sample.
  • the shipment package includes a bar-coded, medium-filled client sample container in an outer container (as discussed herein), a sterile, 60mL syringe, a patient-specific bar-coded shipping container approved for biohazardous materials containing an absorbent sheet, a Tyvek® outer container, sufficient bubble-wrap to stabilize the contents, and foam insulation in an outer corrugated cardboard box (the latter items commercially available from Saf-T-Pak®).
  • the appropriate needles / cannula and other medical supplies are generally accessible equipment which will be supplied by the physician, but may be included as part of the shipment.
  • the method continues by introducing the shipment package components to a processing module of a database via a log-in port by scanning a barcode on the client sample container in the completed recording information.
  • the database is custom-designed to have the ability to comply with the requirements of the American Association of Blood Banks (AABB) standard 6.3 and 21 CFR ⁇ 820.30 (FDA Guidance, January 1 1 , 2002, "General Principles of Software Validation) using, for example, a commercially available program such as Microsoft's Access program.
  • the database includes (but is not limited to), the information obtained from the shipment package to coordinate the client sample with the client; such as the information included in the patient-specific bar-coded shipping container. This information will also be included in a standardized form.
  • the database will be organized in modules similar to the organization in the standardized form, will be searchable, and will be programmed to produce all the various forms associated with this process.
  • one vial each of collagenase, neutral protease, and DNase I is removed from a freezer and thawed in a biosafety cabinet for use in a digestion solution.
  • a pre-defined mixture of collagenase and neutral protease was used, Roche Liberase®, which contains both collagenase I and collagenase II, plus thermolysin, a neutral protease. Thawing at room temperature and without assistance supports in the protection of the integrity and viability of these solutions.
  • the client sample was ' removed from the outer shipping container and gently agitated in the client sample container manually to re-suspend the fat and any sediment in the ⁇ medium, and further, to ensure that the sterility test samples will be representative of the contents.
  • the sample container is wiped with alcohol from spray bottles with filter-sterilized 70% ethanol or iso-propanol to ensure it is not contaminated.
  • the client sample container is elevated or "hung” and allowed to stand undisturbed for approximately five (5) minutes to note the presence of visible blood and estimate the amount of oil from lysed fat as a fraction of the total adipose tissue present in the client sample. This observation is recorded in a defined manner on a pre-designed form. Observations such as the amount of oil present will be entered into a standardized form, and thus the information becomes part of the database. The bar codes on the containers will be scanned, and the information in the barcode will be imported into the database.
  • Sterility of the adipose tissue sample within the client sample container is tested to ensure the quality.
  • the removal, via a bottom port of the client sample container, allows extraction by gravitational force thus eliminating any need for a "pump" etc.
  • Sterility and microbial testing is performed by standard commercial systems such as BacT/ Alert or similar testing. Specific testing procedures are performed in order to comply with and receive required AABB or other professional organization certification and adhere to specific current and future FDA rules as applicable. Sterility samples are kept at room temperature until sent to the
  • the adipose tissue sample is washed by disinfecting one of the top ports of the client sample container, by wiping with 70% or sterile alcohol with a swab, and adding a defined amount of salt solution.
  • the defined amount is at least equal to the volume of adipose tissue sample to wash it effectively.
  • the salt solution e.g. Hank's Balanced Salt Solution (HBSS)
  • HBSS Hank's Balanced Salt Solution
  • the client sample container is gently agitated and allowed to stand undisturbed for a defined time period; about five minutes. Using the same port that was used to obtain the sterility sample, the wash is removed and discarded. The container was allowed to "hang" in an elevated position undisturbed until fat is observed floating in a single layer at the top of the container. Oil dispersed from the adipose tissue sample is removed.
  • the digestion solution prepared is injected into the client sample container having the client/adipose tissue sample to form a digestion mixture within the client sample container.
  • the outside of the vials of the digestion solution was wiped to ensure sterility with an alcohol swab.
  • the digestion solution is injected into the washed adipose tissue sample using one of the top ports of the client sample container.
  • the digestion mixture is incubated at 37 degrees Celsius for 45 minutes while being agitated on a rocking platform at 24 rocks per minute.
  • the adipose tissue is converted from a suspension of tissue fragments up to 4 millimeters in size into a much smoother suspension in which most tissue fragments are less than 1 millimeter in diameter, as most of the adipose tissue is dissociated into isolated mature adipocytes and stromal -vascular fraction cells, although some whitish, connective tissue may remain intact.
  • the solution is centrifuged at a low speed to separate the mature adipocytes from the rest of the digestion mixture.
  • the stromal vascular fraction (“SVF") phase of the centrifuged digestion mixture is withdrawn through a sterile, 40 micrometer mesh filter.
  • the centrifugation of the digestion mixture in the client sample container serves to separate the SVF from the adipocytes and undigested adipose tissue. Removing this SVF from the client sample container to a centrifuge tube and re-centrifuging allows formation of a "tight" pellet at the bottom of the tube (as discussed herein), so that greater than 95 percent, and as much as 99 percent, of the enzyme solution can be removed.
  • the suspension of the filtered digestion mixture is centrifuged in two 50mL tubes upon removal from the client sample container, isolating a "first" stromal vascular pellet.
  • the supernatant of the centrifuged filtered suspension is removed.
  • the stromal vascular "tight" first pellet is re-suspended by trituration in a red blood cell lysis buffer, eliminating red blood cells, as well as removing residual enzymes and debris, forming a cell suspension which is centrifuged to form a "second" pellet.
  • the supernatant of the centrifuged cell suspension is removed.
  • the second pellet is re-suspended by titration adding HBSS forming a "second cell suspension".
  • This second cell suspension is counted and analyzed for viability by using a small aliquot (20 micro liters) of the second cell suspension mixed with an equal volume of a mixture of acridine orange and propidium iodide stains and counted using the Nexcelom Cellometer Vision instrument (Nexcelom Biosciences).
  • the second cell suspension is centrifuged to form a "third" pellet which is stored in a biosafety cabinet for further processing.
  • the "third pellet” defines a stem cell pellet product, e.g. a washed SVF pellet which includes a mixture of cells of pre-adipocytes, adipose-derived mesenchymal stem cells, microvascular endothelial cells, endothelial progenitor cells, monocytes, and small numbers of vascular smooth muscle cells.
  • the mixture must contain no mature adipocytes, and at least 1 % of the nucleated cells in the mixture must be adipose-derived mesenchymal stem cells.
  • the mixture or "stem cell pellet product” or “washed SVF pellet” must exhibit a combined viability by acridine orange/ propidium iodide or trypan blue dye-exclusion assay of no less than 35%. Further, the adipose-derived mesenchymal stem cells contained therein must be capable of proliferation when placed in contact with a suitable culture medium under appropriate environmental conditions known to those skilled in the art of cell culture.
  • the proprietary culture forms are designed to allow the direct flow of information into a computer system/database (as discussed herein).
  • the culture forms include labels that are proprietary and include (but are not limited to) a Client Number (barcoded) and Collection Date (barcoded).
  • the information on the forms is transferred to an electron format, in thi s example, Microsoft Excel®, and thereafter transferred to a data standard for inclusion into a database. Therefore the information is accessible to both the primary facility and the cosmetic product processing facility.
  • the T25 culture flask w'th a minimum of l .Oxl O 6 SVF for shipment to cosmetic product processing facility include a medium 0.5% FBS adipose stem cell medium with ⁇ g/mL Gentamicin as described.
  • 1/2 feed is generally defined as removal of about half the volume of spent medium from flask, then add equal or a little more fresh medium. In the present example, complete medium changes are performed prior to the weekend to ensure nutrient avail ability for cell viability and growth.
  • the T25 culture flask Upon recognition that the T25 culture flask is about 30% to 50% confluent, it is shipped to the cosmetic product processing faci lity (CPPF). To maintain viability of cells, the cells are sent overnight. It is recognized the SVF should not be sent on Friday or the day before a holiday. A copy of the culture record, original processing request, and 6 barcoded labels are included with the shipment to the cosmetic product processing facility. To ensure viability of the culture, it should be shipped in 13 to 15 days from the date of plating. Failure to ship within this time period will require review by a supervisor, management and cosmetic product processing facility. As will be appreciated, the system as described herein will alert laboratory staff of any delayed shipment to ensure the quality and viability of the culture.
  • CPPF cosmetic product processing faci lity
  • the T25 Flask Upon recei pt at the cosmetic product processing facility, ( 1 3 to 1 5 Days post processing) the T25 Flask is "checked in” via the system "connected” to the system at the primary facility.
  • the m edi a in the received T25 F l ask i s reduced 80 to 90 p ercent u pon recei pt and i ncub ated for 1 to 3 day s unti l con fl uence .
  • the contents of the T25 Flask is plated into four (4) T75 flasks coated with human plasma Fibronectin using all recovered cells (a minimum of 300,000 cells per T75 flask). In the present example, no cells will be set aside for storage at this recovery.
  • the T75 Flasks are placed in incubator and fed every 24 to 48 hours as necessary until greater than 80% confluent exists in each flask.
  • the growth media is replaced with non-growth media, HAM's F-12.
  • non-growth media can be limited to a combination of sugars and carbohydrates.
  • the T75 Flasks are incubated for 4 to 7 days allowing secretion of substances, e.g. cytokines, growth factors and proteins, into the non-growth media to create a "conditioned media".
  • Table 1 and replication Tables 1 A and IB illustrate components of a conditioned media created by the method of the present example.
  • Various assays illustrate levels of substances found in the conditioned media using various cell growth medium.
  • the conditioned media is harvested as appropriate and combined with a formulation created recognizi ng 1 ) a use for a particular product, e.g. a face cream, eye cream, skin cream, toner, hair product, serum and/or moisturizer, and 2) appreciating the properties (biological, physical and chemical) of the conditioned media to establish quality as well as effectiveness in use; or "elegance" as recognized by one skilled in the art of blending.
  • Table 2 is the formulation for a serum product
  • Table 3 is the formulation to form an eye cream product
  • Table 4 is the formulation used in a moisturizer product.
  • Each cosmetic formulation has (i) a ph of between 6.0 and 8.0, and (ii) contains no proteases and (iii) is sterile and further, the growth media is either a commercially available media or a specie specific media.
  • each formulation in the present example is designed, based on the principles discussed herein. Specifically, to complement the amount of IL-8 achieved by the process of the present example, and specifically, the "ACC" media, the percentages of the "ingredients" in the formulation would be based on the amount of components, cytokines, proteins and growth factors, as illustrated by TABLES 1 , 1 A and I B found in the "conditioned media".
  • the ACC media which is the subject matter of U.S. Patent No 7,989,205 (American Cryostem Corporation) correlates to an increased product of IL-8 compared to conditioned media obtained by use of the "MC" media product; MesenCult®.
  • the desired elegance of the cosmetic product will include properties such as consistency which will prohibit "running” or “dripping" of the product.
  • thickeners such as carbomers, hyaluronic acid, glycerol, and dimethicone can be used.
  • Another property required is the ability to maintain shelf life of a product for commercial purposes.
  • refrigeration may be required and is acceptable based on the product formulation for a particular product.
  • One skilled in the art would recognize that the ability to have a synergistic effect of the components (including essential cytokines) based a defined formulation would be extremely advantageous for both biological properties of the particular product and commercial brand.
  • the T25 flask for subculture at the pri mary facility; 19 to 24 days post processi ng has an objective to ultimately freeze 5 to 10 PI vials with a minimum of 1 million cells each for retrieval by the cosmetic product processing facility for future products. Priority is to plate 2 T75 flasks as follows:
  • Cells are plated in 0.5% FBS ASC Medium with 101. l .g/mL Gentamicin which will be removed at the first feeding. If BacT " results are negative prior to this, it will be removed immediately. About 2 days prior to expected harvesting while cells are actively growing, feed with 2% HSA Medium.
  • APD population doubling
  • ski l l in the art would recognize quality control recovery is preferable. Specifically, in the current example, recover 1 quality control vial . Perform BacT test on 1 /2 of the final cell suspension after cell count is removed and plate 1 T25 or T 12.5 flask (according to count) coated with Fibronectin using 0.5% FBS ASC Medium . Subculture 2 times after recovery seeding each new flask with Ix l O 4 ASC and harvest for count only after second subculture. If culture is not ready for subculture (or harvest) 14 days after plating, subculture is still performed. If the culture did not perform a complete 3 ⁇ 4 population doubling, it is considered dead and has failed growth QC standard. If the culture did double, then continue with the QC process. Calculate the change in population doubling (APD), and the cumulative population doubling level (cPDL). Do not forget to include the cPDL from the PI freeze:
  • APD Logi c , [Yield / Seed] x 3.32

Abstract

L'invention concerne des procédés, des procédures et des systèmes commerciaux pour la collecte, le stockage et la distribution d'un échantillon biologique pour la production d'un produit de formulation cosmétique.
PCT/US2012/030774 2011-03-28 2012-03-27 Procédés, procédures et systèmes commerciaux pour la collecte, le stockage cryogénique et la distribution de formules cosmétiques à partir d'une matière biologique obtenue à base de cellules souches WO2012135237A2 (fr)

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US14/008,024 US11389394B2 (en) 2011-03-28 2012-03-27 Business methods, processes and systems for collection, cryogenic storage and distribution of cosmetic formulations from an obtained stem cell based biological material
JP2014502724A JP2014518846A (ja) 2011-03-28 2012-03-27 得られた幹細胞ベースの生物学的材料からの美容剤用配合物の収集、低温保管、及び分配のためのビジネスモデル、方法、及びシステム
CN201280026116.1A CN104662532B (zh) 2011-03-28 2012-03-27 基于干细胞的化妆配制品及制备其的方法和系统
HK15110032.2A HK1209503A1 (en) 2011-03-28 2015-10-14 Business methods, processes and systems for collection, cryogenic storage and distribution of cosmetic formulations from an obtained stem cell based biological material
US17/844,198 US20220323342A1 (en) 2011-03-28 2022-06-20 Business methods, processes and systems for collection, cryogenic storage and distribution of cosmetic formulations from an obtained stem cell based biological material

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US201161468132P 2011-03-28 2011-03-28
US61/468,132 2011-03-28
US201161582026P 2011-12-30 2011-12-30
US61/582,026 2011-12-30
US201261588841P 2012-01-20 2012-01-20
US61/588,841 2012-01-20

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US17/844,198 Continuation US20220323342A1 (en) 2011-03-28 2022-06-20 Business methods, processes and systems for collection, cryogenic storage and distribution of cosmetic formulations from an obtained stem cell based biological material

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JP2018065847A (ja) 2018-04-26
US20140140950A1 (en) 2014-05-22
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CN104662532A (zh) 2015-05-27
JP6835891B2 (ja) 2021-02-24
US11389394B2 (en) 2022-07-19
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CN104662532B (zh) 2018-09-07
WO2012135237A3 (fr) 2014-05-01

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